Biomedical Engineering Reference
In-Depth Information
TABL E 4.5.
Identified Sites of Deamidation and Characterization after Incubation in Human Plasma
Identified Sites of Deamidation
Characterization
Native IEF
Band 1/Band 2
OD Ratio
F protein
Binding
ELISA (%)
a
F protein Binding
Kinetics (SPR)
(k
off
)(s
1
)
b
Asn-289
(%)
Asn-318
(%)
Asn-387
(%)
Total
Deamidation (%)
RSV
Microneutralization
Samples
0
3.9
14.0
17.9
3.4
4.96
10
6
Initial
1.6
4.6
19.2
25.4
2.1
47
Neutralizes RSV
6.36
10
6
1 week
0
4.5
24.9
29.4
1.6
88
Neutralizes RSV
5.57
10
6
2 weeks
2.8
4.0
34.5
41.3
1.2
85
Neutralizes RSV
6.07
10
6
3 weeks
2.0
4.3
40.1
46.4
0.75
91
Neutralizes RSV
5.87
10
6
4 weeks
4.8
3.6
47.2
55.6
0.40
52
Neutralizes RSV
5.81
10
6
5 weeks
12.1
4.8
60.3
77.2
0.25
77
Neutralizes RSV
Note: 0, 3.9, 14.0, and 17.9 are reference standards.
a
Value listed is % binding relative to the reference standard.
b
Samples extracted from human plasma had low total protein concentrations. Because the SPR off-rate is independent of protein concentration, it was used to analyze these
samples.
